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Generation of CAR-T Cells by Lentiviral Transduction

Part of the Methods in Molecular Biology book series (MIMB,volume 2312)


CAR-T cell therapy is one of the most successful cell-based therapies. T cells are the most common cells to be genetically modified for cancer therapy, not only because T cells have cytotoxicity but also because they are easily cultured ex vivo and genetically modified with viral vectors. Hence, for nonexperts, T cell engineering is an ideal starting point for mammalian cell engineering or for development of therapeutics. Here, we have described a basic procedure for lentiviral transduction of human primary T cells to generate a CAR-T cell and assays to confirm CAR expression and function.

Key words

  • CAR-T cell
  • Lentivirus vector
  • Dynabeads
  • PEG precipitation
  • Retronectin
  • Flow cytometry
  • CAR-T cell activation assay

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  • DOI: 10.1007/978-1-0716-1441-9_1
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I thank Wilson W. Wong (Boston University) and Jang Hwan Cho (Broad Institute, MIT) for kindly sharing their T cell transduction procedure and accepting to use the data in this paper. I also thank Satoshi Mimura (Hitachi Ltd.) for helping to optimize this procedure. This work was supported by Leading Initiative for Excellent Young Researchers grant from Japan Society for the Promotion of Science.

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Correspondence to Atsushi Okuma .

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Okuma, A. (2021). Generation of CAR-T Cells by Lentiviral Transduction. In: Kojima, R. (eds) Mammalian Cell Engineering. Methods in Molecular Biology, vol 2312. Humana, New York, NY.

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1440-2

  • Online ISBN: 978-1-0716-1441-9

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